Shank 3 Mouse Model of Autism - Psychogenics...Shank 3 Mouse Model of Autism Confidential 2...

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1 Confidential Shank 3 Mouse Model of Autism

Transcript of Shank 3 Mouse Model of Autism - Psychogenics...Shank 3 Mouse Model of Autism Confidential 2...

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    Shank 3 Mouse Model of Autism

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    Background

    • About 2% of people with autism carry harmful mutations in SHANK3, a protein that helps organize the connections between neurons

    • Developed by Guoping Feng, Shank3 –Feng mice harbor a deletion of exons 13-16 of the PDZ domains leading to the deletion of the Shank3α and Shank3β isoforms and partial deletion of Shank3γ

    • These mice exhibit some social, communicative, repetitive, and sensory processing abnormalities associated with autism spectrum disorder.

    • Displays some social deficits• Shows a decrease in activity and locomotion compared to littermate controls,

    including reduced rearing• Shows gait differences compared to littermate controls• Displays some anxiety-like behaviors• Shows reduced startle responses and increased prepulse inhibition of startle

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    No Significant differences in Body Weight Between WT and KO Mice

    Age (weeks)

    5 6 7 8 9 10 11

    Bo

    dy W

    eig

    ht

    (g)

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    SHNAK3/F_WTSHNAK3/F_KO

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    Behavioral Assessments

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    KO Mice Show Deficits in Rotarod PerformanceL

    ate

    nc

    y t

    o F

    all

    (s

    ec

    )

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    Shank3/F_WTShank3/F_KO

    *

    Sp

    eed

    at

    Fall

    (rp

    m)

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    Shank3/F_WTShank3/F_KO

    *

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    Urine Open Field Test

    • Male mice are placed in an Open Field where they are presented with the urine of an estrus female

    • Activity, proximity to the urine, ultrasonic vocalizations, and urine marking by the male are recorded to determine social and function

    • Urine spots developed with Nindyrin Spary

    Estrous female urine spot

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    KO Mice Show Decreased Activity in the Open Field Following Exposure to Female Urine

    To

    tal

    Dis

    tan

    ce T

    ravele

    d (

    cm

    )

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    SHNK3/F_WTSHNK3/F_KO

    *

    Dis

    tan

    ce

    Tra

    ve

    led

    in

    Ce

    nte

    r (c

    m)

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    SHNK3/F_WTSHNK3/F_KO

    *%

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    e S

    pen

    t in

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    nte

    r

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    Reciprocal Social Interaction

    • Same-genotype, -sex, -age dyads are allowed to freely interact for 10 minutes

    • Distance between mice, proximity, and interactions are measures of sociality

    Front Back Side

    Nu

    mb

    er

    of

    Inte

    racti

    on

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    * *

    Tim

    e S

    pe

    nt

    Fo

    llo

    win

    g (

    sec)

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    KO Mice Show Increased Grooming

    Gro

    om

    ing

    Bo

    uts

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    KO Mice Show Decreased Startle Response

    Me

    an

    Sta

    rtle

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    sp

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    KO Mice Show Increased Hindlimb Clasping

    Mice are lifted gently by the tail with front limbs just above the surface

    Clasping of hind legs is noted (normal is a spread in the hind legs)

    Pe

    rcen

    t M

    ice

    Cla

    sp

    ing

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    KO Mice Show Decreased Ultrasonic Vocalizations

    Ult

    ras

    on

    ic V

    oca

    liza

    tio

    ns

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    Shank3/F_WTShank3/F_KO

    *

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    Electrophysiology

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    Impaired corticostriatal synaptic transmission in Shank3 KO

    • Extracellular field potential responses (A) and whole-cell currents in medium spiny neurons (B) were evoked in dorsal striatum by

    stimulating corpus callosum.

    • Brain slices were prepared from male mice aged 10 weeks for extracellular field potential recordings (A) and 14 weeks for whole-

    cell patch clamp recordings (B).

    • Numbers in parenthesis show number of slices (cells); animals used.

    A B

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    Impaired NMDA-mediated synaptic transmission in dorsal striatum in Shank3 KO

    • Whole-cell NMDA-mediated currents in medium spiny neurons were evoked in dorsal striatum by stimulating corpus

    callosum in 14 weeks old male mice.

    • Numbers in parenthesis show number of cells; animals used.

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    Impaired synaptic transmission in nucleus accumbens in Shank3 KO

    • Extracellular field potential responses were evoked in nucleus accumbens (NAc) by stimulating terminals within the

    nucleus in 10 weeks old male mice.

    NAcacp

    Str

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    Molecular markers

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    Decreased expression of synaptic proteins in striatum and BNDF in cortex in Shank3 KO

    • Expression of synaptic proteins in striatum and BDNF in cortex was assessed by qPCR in 16 weeks old male

    Shank3 KO mice (n=10).